Test for Carbohydrates Identifies/ Visible Results Reagents

Used to differentiate… [Compound responsible]

Molisch’s General test for +: Purple ring @ liq. junction  5% Alpha naphthol in
carbohydrates [furfural + alpha naphthol] alcohol
 Sulfuric acid
Anthrone Test General test also +: Deep green Anthrone reagent
[furfural-anthrone complex]
Iodine Test Complex* v.s. Simple Starch: Deep blue  I2
carbs. [Amylose-iodine complex]  KI
Red if Dextrin
*detects HELICAL Brown if Glycogen
Amylose
Seliwanoff's KETOSE (Fructose) v.s. Fructose: Cherry-red RESorcinol in 50% HCl
Aldose [furfural + resorcinol]
Aldose: Blue-green to peach
Or colorless to faint pink
Tauber’s KETOSE v.s. Aldose Bright red Aminoguanidine
Aminoguanidine Test Or Bright reddish purple (Tauber 1949) Sulfuric acid
PENTOSE ID Tests
Bial's PENTOSE v.s. Hexose Pentose: Blue green [furfural  Conc. HCl
reacting with colored complex]  Bial’s reagent
Hexose: Muddy brown-grey  Orcinol +
 FeCl3 +
 Ethanol
Tauber’s PENTOSE +: Cherry red Benzidine
Benzidine Test - More specific,
up to 0.01 mg
Tollen’s PENTOSE +: Betadine-like color Phloroglucinol
Phloroglucinol Glucose (Hexose): Red wine
Reaction Fructose (Hexose): Greenish-
brown
Arabinose: Reddish brown
Test for REDUCING SUGARS (with free aldehyde or ketone group):
Tollen’s Silver Reducing sugars +: Silver mirror Silver nitrate
Mirror Test
Fehling’s Reducing sugars Aldehydes: Brick red PPT  A: CuSO4·5H2O
- ALKALINE pH [Blue Cu (II) reduced to  B: KOH + Rochelle salt
- NOT specific for red Cu (I) - Cuprous oxide] (Sodium potassium
aldehydes tartrate)
Benedict’s Reducing sugars +: Brick red PPT Benedict’s Solution:
- Better than  Sodium citrate
Fehling’s (Na3C6H5O7)+
- Urine exam for  Sodium carbonate +
pathological  CuSO4·5H2O
amt of sugar
Barfoed's Reducing +: Brick red PPT [Cuprous oxide] Barfoed’s reagent
- ACIDIC pH MONOSACCHARIDES Mono: 2-3 mins  Copper (II) acetate +
- NOT for urine Di: after 10 mins  Glacial acetic acid
exam (Cl forms Mono- v.s. Di- Nonreducing: No reaction
green ppt)

VVVELEZ Feb2019
 Trommer’s MONOSACCHARIDES +: Bright red color KOH + CuSO4
Nylander’s Reducing sugars +: Black ppt [metallic Bismuth]  Bismuth nitrate
 Rochelle Salt
 KOH
Moore’s Reducing sugars EXCEPT +: Yellow/ Orange/ Dark brown, Conc. NaOH
SUCROSE Liberating caramel odor
[liberation of aldehydes]
CONFIRMATORY Tests for Reducing Sugars:
Osazone Test ID of Reducing sugar +: Osazone crystals 1:1 Phenylhydrazine and
(Kowarsky Test) MANNOSE (bis-phenylhydrazone) NaOAc

Glucose/Fructose/Mannose:
Needle-shaped
Maltose: Petal / Sunflower
Galactose: Rhombic
Lactose: Powder-puff/ Fluffy ball
Mucic Acid Test GALACTOSE +: Crystals Nitric acid
[Insoluble mucic acid or
galactosaccharic acid]

Test for Proteins Positive for/ Detects Visible Result Reagent/s

Biuret Test General test for Proteins Proteins: Deep violet color  Dilute CuSO4
/Bayuret/ = Violet [presence of peptide (CONH)  NaOH (alkaline soln.)
bond]  Biuret
(H2NCONHCONH2)

 Biuret
complex

If Short peptides: Blue to pink

Ninhydrin Test Free amino and carboxylic Proline (P): Yellow Ninhydrin
- Delicate test acid group in Proteins and AA with free amino group:
Peptides Ruhemann’s Purple

Sakaguchi Test Arginine (R): Guanidine Red  Alpha naphthol

Ms. Argonza likes  Sodium
Japanese men :P hypobromite/chlorite
(oxidizing agent)
Sulfur Test/ Sulfur containing AA: Black PPT [PbS]  40% NaOH
Lead Sulfide Test Methionine (M)  Pb(CH3COO)2 = Lead
Cysteine (C) acetate
Cystine
Sullivan’s Test Cysteine (C) Red  Sodium 1,2 –
napthoquinone-4-
sulfonate
 Sodium hydrosulfite

VVVELEZ Feb2019
Nitroprusside Test Cysteine (C): Sulfhydryl Red complex Nitroprusside in
Alkaline soln.
Xanthoproteic Test Aromatic AA, but NOT Yellow, turns Orange with Conc. Nitric acid
Phenylalanine (F): NaOH
Tyrosine (Y)
Tryptophan (W)
Glycine (G)
and Albumin
Pauly’s Diazo Test Histidine (H) Red [azo dye] Diazotized sulfanilic acid
Tyrosine (Y) in alkaline soln.
Millon’s Test Tyrosine (Y): Phenol group Red or Flesh PPT after heating Mercury nitrate + nitrite
[Mercury phenolate]
Folin’s Test Tyrosine (Y) Blue Alkaline
phosphomolybdotungstic
acid
Folin’s McCarthy Proline (P) Proline + Isatin reagent: Blue
Sullivan Test Methionine (M) Methionine + Sodium
nitroprusside: Red
Hopkins-Cole Test Tryptophan (W): Indole Violet ring  Glyoxylic acid
- Glyoxylic Acid  Conc. H2SO4 (strong
Reaction acid)
Ehrlich Test Tryptophan (W) Blue  p-dimethylamino
benzaldehyde in 12 N
HCl
 10% H2SO4
 0.5% NaNO2
Liebermann’s Test Tryptophan (W) Violet  Conc. HCl
 Sucrose soln.
Heller’s Ring Test  Coagulation Test for White PPT of coagulated protein Conc. HNO3 dropwise
Proteins
 Presence of ALBUMIN
in urine

Precipitation Titration Methods

Notes
Mohr’s Volhard’s Fajan’s
Best used if Neutral solution Acidic medium
Titrant Silver nitrate Silver nitrate
Standard Ammonium thiocyanate Silver nitrate (1° Std)
NH4SCN
(or KSCN)
Modification Add CaCO3 and NaHCO3 Coldwell’s method:
if acidic Add Nitrobenzene or Dibutyl phthalate
to coat Ag ions: protects against
interaction w/ thiocyanate
Indicator Potassium chromate Ferric ammonium sulfate Fluorescein
K2CrO4 Fe(NH4)(SO4) (adsorption indicator)
Endpoint Red Red or reddish brown Green to Pink
Silver chromate Ferric thiocyanate complex
Ag2CrO4 Fe(SCN)3

VVVELEZ Feb2019